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Homogenization of Whole Rat (R. Norvegicus) Lung

Instrument: Precellys® 24 Dual and IKA Ultra-Turrax® T-10

Sample: Whole rat (Rattus norvegicus) lung

Protocol:

  • For the Precellys, place one whole rat lung per tube into 7ml tubes from the hard tissue homogenizing kits.
  • Add 3 volumes per mass of rat lung of 50/50 v/v acetonitrile / 0.9% NaCl solution*.
  • Run Precellys at 6500rpm, 2 x 20 seconds
  • For the Ultra-Turrax, place one rat lung into a 15 ml tube.
  • Add 3 volumes per mass of rat lung of 50/50 v/v acetonitrile / 0.9% NaCl solution*.
  • Run for about 40 seconds.

*The lab performing this protocol was looking to extract previously dosed pharmaceutical compounds.  You may substitute a more appropriate buffer for your application.

Results:

After homogenization, centrifugation, and protein precipitation, 10µl of supernatant was injected into an API3000 triple quadrupole LC/MS/MS system.  The analyte levels in the sample are similar (see the figure below).  Reproducibility is reported to be good in both cases.

Concentration of pharmaceutical compound extracted by processing with Precellys and Ultra-Turrax homogenizers.

Customer reported a Few residual cartilaginous fragments (probably bronchus/trachea) were observed in both methods, but somewhat fewer amounts with the Ultra-Turrax.  The results show
that there is no impact on the analyte recovery from matrix due to the fragments.

With both methods, temperature within the sample after processing could reach 70˚C depending on the sample and protocols.  Considering the stability of the analyte and the short time of exposure to this temperature (sample were preserved in ice before and after processing), sample degradation was reported to be insignificant.